Treatment of autism and autism spectrum disorders with biotin compositions

ABSTRACT

The present application relates to the treatment of autism and autism spectrum disorders using biotin compositions.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. Provisional Application No16/560,826 filed Sep. 4, 2019, which claims priority to of U.S.Provisional Application No 62/727,861 filed Sep. 6, 2018, the contentsof which is incorporated by reference herein in its entirety.

BACKGROUND Field

The present application relates to the treatment of autism and autismspectrum disorders with biotin compositions.

Description of the Related Art

Biotin is an essential water-soluble vitamin also known as Vitamin H,Coenzyme R, and Vitamin B7. It is an essential co-factor for five knowncarboxylases involved in fatty acid biosynthesis, gluconeogenesis,branched-chain amino acid metabolism, fatty acid metabolism,tricarboxylic acid cycle anaplerosis, and pleiotropic gene regulation,particularly for genes in carbohydrate metabolism. Magnesium biotinateis more water soluble than biotin.

Autism, or autism spectrum disorder, is used to refer to a range ofconditions characterized by challenges with social skills, repetitivebehaviors, speech and nonverbal communication. Individuals with anautism spectrum disorder are usually diagnosed between 2 and 3 years ofage. In some cases, it can be diagnosed as early as 18 months. Somedevelopmental delays associated with autism can be identified andaddressed even earlier. Some metabolic disorders have been associatedwith autism.

SUMMARY OF THE INVENTION

In some embodiments, biotin will be used to treat an autism spectrumdisorder. In some embodiments, a biotin salt will be used to treat anautism spectrum disorder. In some embodiments magnesium biotinate willbe used to treat an autism spectrum disorder. In some embodiments, thetype of autism spectrum disorder to be treated may differ. In someembodiments, the type of autism spectrum disorder to be treated may berelated to low biotin levels, impaired carboxylase function, increasedpropionic acid levels, or increased levels of isoleucine, methionine,threonine, and/or valine. In some embodiments, the amount of biotin orbiotin salt administered is between 10mg/day to 10000mg/day. In someembodiments, the amount of biotin or biotin salt administered is between100mg/day and 1000mg/day.In some embodiments, the biotin, biotin salt,or magnesium biotinate will be administered once a day. In someembodiments, the biotin, biotin salt, or magnesium biotinate will beadministered more than once a day. In some embodiments, the biotin,biotin salt, or magnesium biotinate will be administered via an oralroute. In some embodiments, the biotin, biotin salt, or magnesiumbiotinate will be administered via an intraperitoneal route. In someembodiments, the biotin, biotin salt, or magnesium biotinate will beadministered via a transdermal, rectal, or sublingual route. In someembodiments, the amount of magnesium biotinate administered is between10mg/day to 1000mg/day. In some embodiments, the amount of magnesiumbiotinate administered is between 10mg/day and 100mg/day. In someembodiments, the biotin, biotin salt, or magnesium biotinate can beprovided as a drug, supplement, medical food, food or biologic. In someembodiments, the biotin, biotin salt, or magnesium biotinate isadministered alone. In some embodiments, the biotin, biotin salt, ormagnesium biotinate is administered in combination with anothertreatment. In some embodiments, the biotin, biotin salt, or magnesiumbiotinate is administered for 1 or more days. In some embodiments, thebiotin, biotin salt, or magnesium biotinate is administered for 1 ormore weeks. In some embodiments, the biotin, biotin salt, or magnesiumbiotinate is administered for 1 or more months. In some embodiments, thebiotin, biotin salt, or magnesium biotinate is administered for 1 ormore years. In some embodiments, the biotin, biotin salt, or magnesiumbiotinate is administered to a pregnant woman or to a woman attemptingto become pregnant to prevent a child from developing autism. In someembodiments, the amount of biotin, biotin salt, or magnesium biotinateadministered to the pregnant woman may be the same throughout thepregnancy. In some embodiments, the amount of biotin, biotin salt, ormagnesium biotinate administered to the pregnant woman may differ ineach trimester. In some embodiments, the amount of biotin, biotin salt,or magnesium biotinate administered to the pregnant woman may be higherduring the second trimester.

In some embodiments, the invention provides a method of treatmentcomprising assessing the biotin status of an individual with an autismspectrum disorder and subsequently providing biotin, biotin salt, ormagnesium biotinate to the individual to treat the autism spectrumdisorder. In some embodiments, assessing the biotin status of theindividual with an autism spectrum disorder comprises obtaining samplesfrom the individual and testing for markers associated with biotindeficiency. In some embodiments, the samples are obtained from blood,serum, peripheral blood mononuclear cell (PBMC), saliva, urine, feces orsweat from the individual with an autism spectrum disorder. In someembodiments, the samples can be tested for markers indicative ofimpaired carboxylases. In some embodiments, the samples can be testedfor impaired propionyl-CoA carboxylase (PCC) or pyruvate carboxylase(PC). In some embodiments, the samples can be tested for amino acidsfound at abnormal levels due to impaired PCC or PC or other carboxylase.In some embodiments, the samples can be tested for biotin, propionicacid, pyruvate, or lactate. In some embodiments, the samples can betested for isoleucine, methionine, threonine, and valine. In someembodiments, the samples can be tested for ratios of one amino acid overanother. In some embodiments, the samples can be tested for white bloodcell count. In some embodiments, the samples can be tested formitochondrial function.

DETAILED DESCRIPTION

The terminology used in the description presented herein is not intendedto be interpreted in any limited or restrictive manner, simply becauseit is being utilized in conjunction with a detailed description ofcertain specific embodiments described herein. Furthermore, embodimentsdescribed herein can include several novel features, no single one ofwhich is solely responsible for its desirable attributes or which isessential to practicing the embodiments described herein.

As used herein, “identifying,” refers to detecting or selecting asubject from a population of potential subjects, for example, toestablish that a particular subject possesses certain properties orcharacteristics. “Identifying” may include, for example,self-identification, self-diagnosis, and diagnosis by a medicalprofessional.

As used herein, “treat,” “treatment,” or “treating,” refers toadministering or providing a composition for prophylactic and/ortherapeutic purposes.

As used herein, the terms “prophylactic treatment,” “prevent,” or“preventing,” can refer to treating a subject who does not yet exhibitsymptoms of a disease or condition, but who is susceptible to, orotherwise at risk of, a particular disease or condition, whereby thetreatment reduces the likelihood that the subject will develop thedisease or condition. A “disorder” is any condition that would benefitfrom treatment with the compositions described herein.

The term “biotin” means D-biotin, an essential water-soluble vitaminalso known as Vitamin H, Coenzyme R, or vitamin B7. D-Biotin hasChemical Abstracts Service Registry No. 58-85-5 and the general formula:

As used herein, the term “biotin salt” refers to an organic, orinorganic salt of D-biotin. Typical salts include alkali metal, alkalineearth metal, ammonia, or organic amine salts as, for example, sodium,potassium, magnesium, calcium, protonated amines such as those derivedfrom ethylamine, triethylamine, ethanolamine, diethylamino-ethanol,ethylenediamine, piperidine, morpholine, 2-piperidinoethanol,benzylamine, procaine and the like

As used herein, the term “magnesium biotinate” refers to the magnesiumsalt of D-biotin, including magnesium hemi-biotinate. MagnesiumD-biotinate is the magnesium salt of the carboxylic acid D-biotin, anddoes not occur naturally. In some embodiments, magnesium D-biotinate isa stable, non-hygroscopic, off-white powder having a definedcomposition, a molecular formula of Mg(C₁₀H₁₅N₂O₃S)₂ and a generalformula of

Some embodiments provide physiologically compatible magnesium biotinatehydrates, crystalline forms, polymorphic forms, solid forms havingspecific bulk densities or tap densities, and solid forms havingspecific particle sizes. Some embodiments provide compositions coatedwith pharmaceutically acceptable materials intended to modify itsrelease and/or bioavailability, including, but not limited to Eudragit,microcrystalline cellulose, hydroxypropylmethylcellulose phthalate, andthe like.

As used herein, the term “magnesium” refers to the magnesium ion, Mg²⁺.

As used herein, the term “pharmaceutically acceptable solvent” can referto water, water for injection, aqueous buffer solutions that arephysiologically compatible, or aqueous solutions containing organicsolvents that are physiologically compatible. A non-comprehensive listof pharmaceutically acceptable solvents is provided in U.S. Departmentof Health & Human Services, Food & Drug Administration, “Guidance forIndustry: Q3C Impurities: Residual Solvents,” December 1997 or itscurrent issue.

As used herein, the term “bioavailability” refers to the amount of asubstance that is absorbed in the intestines and ultimately availablefor biological activity in a subject's tissue and cells.

As used herein, the term “excipient material” refers to any compoundthat is part of a formulation that is not an active ingredient, i.e.,one that has no relevant biological activity, and which is added to theformulation to provide specific characteristics to the dosage form,including by way of example, providing protection to the activeingredient from chemical degradation, facilitating release of a tabletor caplet from equipment in which it is formed, and so forth.

For oral administration, the compositions disclosed herein can beprovided as a tablet, aqueous or oil suspension, dispersible powder orgranule, emulsion, hard or soft capsule, syrup, elixir, or beverage.Solid dosage forms such as tablets and capsules may comprise an entericcoating. Compositions intended for oral use can be prepared according toany method known in the art for the manufacture of pharmaceuticallyacceptable compositions and such compositions may include one or more ofthe following agents: sweeteners, flavoring agents, coloring agents,coatings, and preservatives. The sweetening and flavoring agents willincrease the palatability of the preparation. Tablets containing thecomplexes in admixture with non-toxic pharmaceutically acceptableexcipients suitable for tablet manufacture are acceptable.

Pharmaceutically acceptable vehicles such as excipients are compatiblewith the other ingredients of the formulation (as well as non-injuriousto the patient). Such excipients include inert diluents such as calciumcarbonate, sodium carbonate, lactose, calcium phosphate or sodiumphosphate; granulating and disintegrating agents, such as corn starch oralginic acid; binding agents such as starch, gelatin or acacia; andlubricating agents such as magnesium stearate, stearic acid or talc.Tablets can be uncoated or can be coated by known techniques to delaydisintegration and absorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period of time. For example, atime delay material such as glyceryl monostearate or glyceryl distearatealone or with a wax can be employed.

Formulations for oral use can also be presented as hardgelatin-containing or non-gelatinous capsules wherein the biotin, biotinsalt, or magnesium biotinate is mixed with an inert solid diluent, forexample calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, such as peanut oil, liquid paraffin or olive oil. Aqueoussuspensions can contain the complex of the biotin, biotin salt, ormagnesium biotinate admixed with excipients suitable for the manufactureof aqueous suspensions. Such excipients include suspending agents,dispersing or wetting agents, one or more preservatives, one or morecoloring agents, one or more flavoring agents and one or more sweeteningagents such as sucrose or saccharin.

Oil suspensions can be formulated by suspending the biotin, biotin salt,or magnesium biotinate in a vegetable oil, such as arachis oil, oliveoil, sesame oil or coconut oil, or in a mineral oil such as liquidparaffin. The oil suspension can contain a thickening agent, such asbeeswax, hard paraffin or cetyl alcohol. Sweetening agents, such asthose set forth above, and flavoring agents can be added to provide apalatable oral preparation. These compositions can be preserved by anadded antioxidant such as ascorbic acid. Dispersible powders andgranules suitable for preparation of an aqueous suspension by theaddition of water can provide the biotin, biotin salt, or magnesiumbiotinate in admixture with a dispersing or wetting agent, a suspendingagent, and one or more preservatives. Additional excipients, for examplesweetening, flavoring and coloring agents, can also be present.

Syrups and elixirs can be formulated with sweetening agents, such asglycerol, sorbitol or sucrose. Such formulations can also contain ademulcent, a preservative, a flavoring or a coloring agent.

Compositions for parenteral administration can be in the form of asterile injectable preparation, such as a sterile injectable aqueous oroleaginous suspension. This suspension can be formulated according tomethods well known in the art using suitable dispersing or wettingagents and suspending agents. The sterile injectable preparation canalso be a sterile injectable solution or suspension in a non-toxicparenterally-acceptable diluent or solvent, such as a solution in1,3-butanediol. Suitable diluents include, for example, water, Ringer'ssolution and isotonic sodium chloride solution. In addition, sterilefixed oils can be employed conventionally as a solvent or suspendingmedium. For this purpose, any bland fixed oil can be employed includingsynthetic mono or diglycerides. In addition, fatty acids such as oleicacid can likewise be used in the preparation of injectable preparations.

Aqueous suspensions may contain the biotin, biotin salt, or magnesiumbiotinate in admixture with excipients suitable for the manufacture ofaqueous suspensions. Such excipients include suspending agents,dispersing or wetting agents, one or more preservatives, one or morecoloring agents, one or more flavoring agents and one or more sweeteningagents such as sucrose or saccharin.

Controlled release vehicles are well known to those of skill in thepharmaceutical sciences, and these aspects can be applied to nutritionaland dietary supplements. The technology and products in this art arevariably referred to as controlled release, sustained release, prolongedaction, depot, repository, delayed action, retarded release and timedrelease; the words “controlled release” as used herein is intended toincorporate each of the foregoing technologies.

Numerous controlled release vehicles are known, including biodegradableor bioerodable polymers such as polylactic acid, polyglycolic acid, andregenerated collagen. Known controlled release drug delivery devicesinclude creams, lotions, tablets, capsules, gels, microspheres,liposomes, ocular inserts, minipumps, and other infusion devices such aspumps and syringes. Implantable or injectable polymer matrices, andtransdermal formulations, from which active ingredients are slowlyreleased, are also well known and can be used in the disclosed methods.

Controlled release preparations can be achieved by the use of polymersto form complexes with or absorb the biotin, biotin salt, or magnesiumbiotinate. The controlled delivery can be exercised by selectingappropriate macromolecules such as polyesters, polyamino acids,polyvinylpyrrolidone, ethylenevinyl acetate, methylcellulose,carboxymethylcellulose, and protamine sulfate, and the concentration ofthese macromolecule as well as the methods of incorporation are selectedin order to control release of biotin, biotin salt, or magnesiumbiotinate.

Controlled release of biotin, biotin salt, or magnesium biotinate can betaken to mean any of the extended release dosage forms. The followingterms may be considered to be substantially equivalent to controlledrelease, for the purposes of the present disclosure: continuous release,controlled release, delayed release, depot, gradual release, long termrelease, programmed release, prolonged release, programmed release,proportionate release, protracted release, repository, retard, slowrelease, spaced release, sustained release, time coat, time release,delayed action, extended action, layered time action, long acting,prolonged action, sustained action medications and extended release,release in terms of pH level in the gut and intestine, breakdown of themolecule and based on the absorption and bioavailability.

Hydrogels, wherein biotin, biotin salt, or magnesium biotinate isdissolved in an aqueous constituent to gradually release over time, canbe prepared by copolymerization of hydrophilic mono-olefinic monomerssuch as ethylene glycol methacrylate. Matrix devices, wherein biotin,biotin salt, or magnesium biotinate is dispersed in a matrix of carriermaterial, can be used. The carrier can be porous, non-porous, solid,semi-solid, permeable or impermeable. Alternatively, a device comprisinga central reservoir of magnesium biotinate surrounded by a ratecontrolling membrane can be used to control the release of the complex.Rate controlling membranes include ethylene-vinyl acetate copolymer orbutylene terephthalate/polytetramethylene ether terephthalate. Use ofsilicon rubber or ethylene-vinyl alcohol depots are also contemplated.

Controlled release oral formulations are also well known. In oneembodiment, the active complex is incorporated into a soluble orerodible matrix, such as a pill or a lozenge. In another example, theoral formulations can be a liquid used for sublingual administration.These liquid compositions can also be in the form a gel or a paste.Hydrophilic gums, such as hydroxymethylcellulose, are commonly used. Alubricating agent such as magnesium stearate, stearic acid, or calciumstearate can be used to aid in the tableting process.

Biotin, biotin salt, or magnesium biotinate may also be deliverytopically, including in a salve, cream, lotion, ointment, shampoo,cosmetic, or emulsion.

The compositions may be administered once, twice, three times per day,or more. In some aspects, the compositions are administered four times aday. For example, the compositions may be administered before, after, orduring a meal. Dosing for oral administration may be with a regimencalling for single daily dose, or for a single dose every other day, orfor a single dose within 72 hours of the first administered dose, or formultiple, spaced doses throughout the day. In some embodiments, whereinbiotin, a biotin salt, or magnesium biotinate is combined with anothertreatment in a combination therapy, the biotin, biotin salt, ormagnesium biotinate and the other active agents which make up thecombination therapy may be administered simultaneously, either in acombined dosage form or in separate dosage forms intended forsubstantially simultaneous oral administration. The biotin, biotin salt,or magnesium biotinate and the other active agents which make up thecombination therapy may also be administered sequentially, with eitherthe biotin, biotin salt, or magnesium biotinate and the other activecomponent being administered by a regimen calling for two-stepingestion. Thus, a regimen may call for sequential administration of thebiotin, biotin salt, or magnesium biotinate and the other active agentswith spaced-apart ingestion of the separate compositions. The timeperiod between the multiple ingestion steps may range from a few minutesto as long as about 72 hours, depending upon the properties of eachcomposition such as potency, solubility, bioavailability, plasmahalf-life and kinetic profile of the agent, as well as depending uponthe age and condition of the patient. The compositions of thecombination therapy, i.e., biotin, biotin salt, or magnesium biotinateand the otheractive agents, whether administered simultaneously,substantially simultaneously, or sequentially, may involve a regimencalling for administration of one composition by oral route and theother composition by intravenous route. Whether the compositions of acombined therapy are administered by oral or intravenous route,separately or together, each such composition will be a suitablepharmaceutical formulation of pharmaceutically-acceptable excipients,diluents or other formulations components.

Active ingredients (e.g., biotin, biotin salt, or magnesium biotinateand the other active ingredients of a combination therapy) can beadministered by the oral route in solid dosage forms, such as tablets,capsules, and powders, or in liquid dosage forms, such as elixirs,syrups, and suspensions. The biotin, biotin salt, or magnesium biotinateand the other active ingredients of a combination therapy can beadministered by the parenteral route in liquid dosage forms. Thecomposition can be made in the form of a dosage unit containing aparticular amount of each active ingredient. One example of an oraldosage form of a composition of the present application is an admixtureof powders contained within a sachet. Because a composition of thepresent application is not hygroscopic and has no repugnant taste orodor, the admixture of powders comprising a composition of the presentapplication can be sprinkled on food or stirred into beverages toenhance ease of use and support high levels of compliance with dailydosage regimens.

In general, the dosage forms of compositions of this disclosure can beprepared by conventional techniques, as are described in Remington'sPharmaceutical Sciences, a standard reference in this field [Gennaro AR,Ed. Remington: The Science and Practice of Pharmacy. 20^(th) Edition.Baltimore: Lippincott, Williams & Williams, 2000]. For therapeuticpurposes, the active components of a single, ora combination therapyapplication can be combined with one or more adjuvants appropriate tothe indicated route of administration. The components may be admixedwith lactose, sucrose, starch powder, cellulose esters of alkanoicacids, cellulose alkyl esters, talc, stearic acid, magnesium stearate,gelatin, acacia gum, sodium alginate, polyvinylpyrrolidone, and/orpolyvinyl alcohol, and then tableted or encapsulated for convenientadministration, the amounts of which are ascertainable by the skilledartisan. Such capsules or tablets may contain a controlled-releaseformulation as may be provided in a dispersion of active compound inhydroxypropyl methylcellulose. Solid dosage forms can be manufactured assustained release products to provide for continuous release ofmedication over a period of hours. Compressed tablets can be sugarcoated or film coated to mask any unpleasant taste and protect thetablet from the atmosphere, or enteric coated for selectivedisintegration in the gastrointestinal tract. Both the solid and liquidoral dosage forms can contain coloring and flavoring to increase patientacceptance. Other adjuvants and modes of administration are well andwidely known in the pharmaceutical art and these aspects can also beapplied to any of the nutritional or dietary supplements describedherein.

While the present invention has been described in some detail forpurposes of clarity and understanding, one will appreciate that variouschanges in form and detail can be made without departing from the truescope of the invention.

EXAMPLES Example 1. Treatment of Autism Spectrum Disorder

In a double-blind clinical study, 20 subjects are divided into twogroups (n=10). Inclusion criteria for the subjects include a diagnosisof an autism spectrum disorder, age<18 years old and low biotin serumlevels at study start date. The control group receives a placebo, thetrial group receives magnesium biotinate at 10-100 mg/day for 12 weeks.Biotin serum levels, carboxylase levels from peripheral bloodmononuclear cells, and magnesium serum levels will be measured atbaseline and at week 4, 8 and 12. Intellectual abilities, behavior,quality of life, developmental age, nonverbal intellectual ability andother measures of autism symptoms will be assessed at baseline and atweek 4, 8 and 12. After week 12, both groups will receive the magnesiumbiotinate. Further assessment of intellectual abilities, behavior,quality of life, developmental age, nonverbal intellectual ability andother measures of autism symptoms will be conducted every 4 weeks.

The above description discloses several methods and materials of thepresent invention. This invention is susceptible to modifications in themethods and materials, as well as alterations in the fabrication methodsand equipment. Such modifications will become apparent to those skilledin the art from a consideration of this disclosure or practice of theinvention disclosed herein. Consequently, it is not intended that thisinvention be limited to the specific embodiments disclosed herein, butthat it cover all modifications and alternatives coming within the truescope and spirit of the invention.

When introducing elements of the present application or the preferredembodiment(s) thereof, the articles “a”, “an”, “the” and “said” areintended to mean that there are one or more of the elements. The terms“comprising”, “including” and “having” are intended to be inclusive andmean that there may be additional elements other than the listedelements.

Without further elaboration, it is believed that one skilled in the artcan, using the preceding description, utilize the present disclosure toits fullest extent. The specific embodiments are, therefore, to beconstrued as merely illustrative, and not limitative of the remainder ofthe disclosure in any way whatsoever. While the present disclosure hasbeen described in some detail for purposes of clarity and understanding,one will appreciate that various changes in form and detail can be madewithout departing from the true scope of the application.

All references cited herein, including but not limited to published andunpublished applications, patents, and literature references, areincorporated herein by reference in their entirety and are hereby made apart of this specification. To the extent publications and patents orpatent applications incorporated by reference contradict the disclosurecontained in the specification, the specification is intended tosupersede and/or take precedence over any such contradictory material.

1.-20. (canceled).
 21. A method for increasing carboxylase serum levelsin an individual, the method comprising administering magnesiumbiotinate to the individual.
 22. The method of claim 21, wherein samplesare obtained from blood, serum, peripheral blood mononuclear cell(PBMC), saliva, urine, feces or sweat from the individual.
 23. Themethod of claim 21, wherein the magnesium biotinate is administered atleast once a day.
 24. The method of claim 21, wherein the magnesiumbiotinate is administered by a route selected from the group consistingof oral, intraperitoneal, transdermal, rectal, and sublingual.
 25. Themethod of claim 24, wherein the magnesium biotinate is administered viaan oral route.
 26. The method of claim 25, wherein the magnesiumbiotinate is formulated as a tablet, aqueous or oil suspension,dispersible powder or granule, emulsion, hard or soft capsule, syrup,elixir, or beverage.
 27. The method of claim 21, wherein an amount ofmagnesium biotinate administered is between about 10 mg/day and about1000 mg/day.
 28. The method of claim 27, wherein the amount of magnesiumbiotinate administered is between about 10 mg/day and about 100 mg/day.29. The method of claim 21, wherein the magnesium biotinate is providedas a drug, supplement, food, medical food, or biologic.
 30. The methodof claim 21, wherein the magnesium biotinate is administered alone. 31.The method of claim 21, wherein the magnesium biotinate is administeredwith another compound.
 32. The method of claim 21, wherein the magnesiumbiotinate is administered for at least one day.
 33. The method of claim21, wherein the magnesium biotinate is administered for at least oneweek.
 34. The method of claim 21, wherein the magnesium biotinate isadministered for at least one month.
 35. The method of claim 21, whereinthe magnesium biotinate is administered to a pregnant woman.
 36. Themethod of claim 21, wherein the individual is a child.
 37. The method ofclaim 36, wherein the child has autism spectrum disorder.
 38. The methodof claim 21, wherein after the administration of the magnesiumbiotinate, the intellectual abilities of the individual are improved.39. The method of claim 21, wherein after the administration of themagnesium biotinate, the non-verbal intellectual abilities of theindividual are improved.
 40. The method of claim 21, wherein after theadministration of magnesium biotinate, the development age of theindividual is improved.